We will be collaborating with centers which are generating genetic expression data on both human tumors and normal tissues to seek candidate proteins suitable for immune attack. These will be interrogated using a new mouse platform wherein mice bearing human HLA molecules are vaccinated with candidate proteins from human tumors. Resulting immune responses will be identified by fluorescent reporter genes also engineered into the mice and the reactive T-cells cloned. If they are then confirmed to react with appropriate human tumors, the mouse T-cell receptors can be directly introduced into human T-cells where they have already been shown to function in vitro as well as in vivo (in clincal protocols against melanoma in the Surgery Branch). In addition, we will be studying the inhibitory mechanisms which appear to impede the accumulation and function of tumor-reactive T-cells in non-melanoma tumors, a phenomenon absent in most melanomas (which can be shown to harbor such cells in the majority of cases). For this effort we will be studying human renal cancer because clinical evidence indicates that an immune response is likely to be present as it can respond to a variety of immunotherapies. Yet efforts to clone tumor-reactive T-cells from renal cancer patients have not been very successful. Known immune inhibitory circuits will be examined to determine if they are active in kidney cancer and if they can be shut off, allowing the recovery of tumor-reactive immune cells. Again the clinical translational thrust of this work will be to clone the T-cell receptors and engineer them into the normal lymphocytes of other patients with renal cancer for clinical administration.